DESCRIPTION: (Applicant's abstract) Microelectromechanical Systems (MEMS) are posed to potentially revolutionize DNA sequencing. We propose a stream- lined program to implement a fully automated, miniaturized system for this application. The automated MEMS system will combine the best features of previous single and several-lane electrophoresis systems developed at MIT and as derived from the world- wide development including: Large-area (75mm x 170mm) separation element fabricated in glass and silicon using MEMS technology with 384 lanes (100 Rm wide, 40 Wm deep channels on 200 zm centers) Piezoelectrically-driven sample injection methods Defect free casting of linear polyacrylamide sieving matrix capable of single-base resolution at 400 bases in multi-lane format Detection based on side-illumination coupled to image-intensified CCD readout with interface to base-calling software Key technology components of the program are: A system capable of running 384 lanes in parallel, with a resolution of 400 bases per lane and run times of approximately one hour. Such an instrument operated full time will allow collection of over 3Mbases of raw sequence data per day per machine, or the potential assembly of about 4Mbases (at Sx redundancy) of finished sequence per week. The prototype instrument will be evaluated in the WVMIT Center for Genome Research. The testing entails integration with sample preparation and data handling capabilities at the CGR. We will begin pilot insertions into the sequence production line after 24 months. Further system improvements through advanced drive for improved resolution and throughput, microfabricated inorganic matrices, and scaled silicon systems will be implemented. An Important structural feature is the co-location of the full team of cross disciplinary engineers, biologists and chemists, extensive microfabrication facilities, and an important genetics laboratory, all at a single site.